Prevalence and impact of single-strain starter cultures of lactic acid bacteria on metabolite formation in sourdough.

Flavour of type II sourdoughs is influenced by the ingredients, processing conditions, and starter culture composition. It is, however, not fully clear to what extent different sourdough lactic acid bacteria (LAB) contribute to flavour. Therefore, two types of flour (rye and wheat) and different LAB starter culture strains were used to prepare sourdoughs, thereby leaving the yeast microbiota uncontrolled. All LAB starter culture strains tested were shown to be prevalent and to acidify the flour/water mixture to pH values between 3.1 and 3.9 after 24h of fermentation. Multiple aldehydes, alcohols, ketones, and carboxylic acids were produced by the sourdough-associated microbiota throughout the fermentation period. Based on the organoleptic evaluation of breads produced with these sourdoughs, five LAB strains were selected to perform prolonged wheat and rye fermentations as to their capacity to result in an acidic (Lactobacillus fermentum IMDO 130101, Lactobacillus plantarum IMDO 130201, and Lactobacillus crustorum LMG 23699), buttermilk-like (Lactobacillus amylovorus DCE 471), or fruity flavour (Lactobacillus sakei CG1). Upon prolonged fermentation, higher metabolite concentrations were produced. For instance, L. sakei CG1 produced the highest amounts of 3-methyl-1-butanol, which was further converted into 3-methylbutyl acetate. The latter compound resulted in a fruity banana flavour after 48h of fermentation, probably due to yeast interference. Rye fermentations resulted in sourdoughs richer in volatiles than wheat, including 3-methyl-1-butanol, 2-phenylethanol, and ethyl acetate.

Ravyts F ，De Vuyst L 《-》

Lactic acid bacteria community dynamics and metabolite production of rye sourdough fermentations share characteristics of wheat and spelt sourdough fermentations.

Four spontaneous rye sourdough fermentations were performed over a period of ten days with daily back-slopping. Samples taken at all refreshment steps were used for culture-dependent and culture-independent characterization of the microbiota present. Furthermore, an extensive metabolite target analysis was performed through a combination of various chromatographic methods, including liquid chromatography coupled to mass spectrometry (LC/MS) and gas chromatography coupled to mass spectrometry (GC/MS). Spearman's rank correlation coefficients were calculated and a principal component analysis (PCA) was performed on the data obtained in this study combined with data obtained previously for wheat and spelt sourdoughs. In general, the establishment of a stable microbial ecosystem occurred through a three-phase evolution, with mainly Lactobacillus plantarum and Lactobacillus fermentum dominating the rye sourdough ecosystems. PCA revealed that ornithine and mannitol were positively correlated with rye sourdoughs, contributing to bacterial competitiveness at the onset of sourdough production. Wheat and spelt sourdoughs showed a high degree of similarity, although certain compounds (e.g. indolelactic acid) appeared to be specific for spelt sourdoughs. The production of amino acid metabolites, mainly hydroxy acids (e.g. phenyllactic acid) and alcohols (e.g. 3-methyl-1-butanol), contributed to the equilibration of the redox balance and further enhanced the competitiveness of dominant species in stable sourdoughs.

Weckx S ，Van der Meulen R ，Maes D ，Scheirlinck I ，Huys G ，Vandamme P ，De Vuyst L ... -  《-》

Robustness of Lactobacillus plantarum starters during daily propagation of wheat flour sourdough type I.

This study aimed at investigating the robustness of selected sourdough strains of Lactobacillus plantarum. Seven strains were singly used as sourdough type I starters under daily back-slopping propagation (ten days) using wheat flour. Cell numbers of presumptive lactic acid bacteria varied slightly (median values of 9.13-9.46 log cfu g(-1)) between and within started sourdoughs, as well as the acidifying activity (median values of 1.24-1.33). After three days also the control sourdough (unstarted) had the same values. As shown by RAPD-PCR analysis, five (DB200, 3DM, G10C3, 12H1 and LP20) out of seven strains maintained elevated cell numbers (ca. 9 log cfu g(-1)) throughout ten days. The other two strains progressively decreased to less than 5 log cfu g(-1). As identified by partial sequencing of 16S rRNA and recA genes, L. plantarum (11 isolates), pediococci (7), Lactobacillus casei (3) and Lactobacillus rossiae (2) dominated the flour microbiota. Monitoring of lactic acid bacteria during sourdough propagation was carried out by culture dependent approach and using PCR-DGGE (Denaturing Gradient Gel Electrophoresis). Except for the sourdough started with L. plantarum LP20, in all other sourdoughs at least one autochthonous strain of L. plantarum emerged. All emerging strains of L. plantarum showed different RAPD-PCR profiles. L. rossiae and Pediococcus pentosaceus were only found in the control and sourdough started with strain 12H1. The characterization of the catabolic profiles of sourdoughs (Biolog System) showed that sourdoughs containing persistent starters behaved similarly and their profiles were clearly differentiated from the others. One persistent strain (DB200) of L. plantarum and Lactobacillus sanfranciscensis LS44, previously shown to be persistent (Siragusa et al., 2009), were used as the mixed starter to produce a wheat flour sourdough. Both strains cohabited and dominated during ten days of propagation.

Minervini F ，De Angelis M ，Di Cagno R ，Pinto D ，Siragusa S ，Rizzello CG ，Gobbetti M ... -  《-》

Biodiversity of lactic acid bacteria and yeasts in spontaneously-fermented buckwheat and teff sourdoughs.

In this study, four different laboratory scale gluten-free (GF) sourdoughs were developed from buckwheat or teff flours. The fermentations were initiated by the spontaneous biota of the flours and developed under two technological conditions (A and B). Sourdoughs were propagated by continuous back-slopping until the stability was reached. The composition of the stable biota occurring in each sourdough was assessed using both culture-dependent and -independent techniques. Overall, a broad spectrum of lactic acid bacteria (LAB) and yeasts species, belonging mainly to the genera Lactobacillus, Pediococcus, Leuconostoc, Kazachstania and Candida, were identified in the stable sourdoughs. Buckwheat and teff sourdoughs were dominated mainly by obligate or facultative heterofermentative LAB, which are commonly associated with traditional wheat or rye sourdoughs. However, the spontaneous fermentation of the GF flours resulted also in the selection of species which are not consider endemic to traditional sourdoughs, i.e. Pediococcus pentosaceus, Leuconostoc holzapfelii, Lactobacillus gallinarum, Lactobacillus vaginalis, Lactobacillus sakei, Lactobacillus graminis and Weissella cibaria. In general, the composition of the stable biota was strongly affected by the fermentation conditions, whilst Lactobacillus plantarum dominated in all buckwheat sourdoughs. Lactobacillus pontis is described for the first time as dominant species in teff sourdough. Among yeasts, Saccharomyces cerevisiae and Candida glabrata dominated teff sourdoughs, whereas the solely Kazachstania barnetti was isolated in buckwheat sourdough developed under condition A. This study allowed the identification and isolation of LAB and yeasts species which are highly competitive during fermentation of buckwheat or teff flours. Representatives of these species can be selected as starters for the production of sourdough destined to GF bread production.

Moroni AV ，Arendt EK ，Dal Bello F 《-》

The addition of citrate stimulates the production of acetoin and diacetyl by a citrate-positive Lactobacillus crustorum strain during wheat sourdough fermentation.

In traditional sourdough fermentation recipes of artisan bakeries, often extra ingredients are added to the flour-water mixture. This may accentuate the aroma and taste of the baked products produced from such sourdoughs. This is made possible, for instance, by stimulating certain microbial activities during fermentation. This study examined the effects of the addition of citrate (a food-grade organic acid present in milk and lemons) on wheat sourdough fermentation in the presence of a citrate-positive, homofermentative, lactic acid bacterial starter culture strain, namely Lactobacillus crustorum LMG 23699. Both liquid and firm wheat sourdoughs were produced. The starter culture strain was able to steer all wheat sourdough fermentations performed, as it always prevailed due to its competitiveness, as shown through culture-dependent microbiological plating and culture-independent bacterial community profiling. Moreover, it possessed all enzyme-encoding genes (as unraveled through genome mining) necessary to convert citrate into desirable compounds such as lactic acid, acetic acid, succinate, acetoin, diacetyl, and 2,3-butanediol. Indeed, citrate addition to the wheat flour-water mixture had an impact on the sourdough fermentation dynamics and thus on the aroma profile of the liquid and firm sourdoughs produced and breads made thereof. A higher final pH, higher total titratable acidity values, and low yeast counts were found in wheat sourdoughs produced with citrate. In particular, the starter culture strain added converted the supplemented citrate into more l-lactic acid as well as acetoin and diacetyl (buttery aroma compounds), which was independent of the dough yield. The buttery aroma compounds were also accentuated in the concomitant breads produced. Further, organic acid production was stimulated in the sourdoughs, whereas increased pyrazine concentrations occurred in the breads. Consequently, citrate supplementation to wheat sourdoughs could be of interest to produce baked goods with enhanced buttery aroma compounds and notes.

Comasio A ，Harth H ，Weckx S ，De Vuyst L ... -  《-》